Pharmaceutical composition containing antibody-drug conjugate and medical use thereof

Abstract

The present invention relates to a pharmaceutical composition containing an antibody-drug conjugate and the medical use thereof. In particular, the present invention relates to a pharmaceutical composition comprising an antibody-drug conjugate and a buffer. The pharmaceutical composition of the present invention has good stability.

Description

Antibody-drug conjugates, pharmaceutical compositions and their pharmaceutical uses Technical Field

[0001] This disclosure pertains to the field of pharmaceutical formulations, specifically relating to a pharmaceutical composition comprising an antibody-drug conjugate and its use as an anticancer drug. Background Technology

[0002] The statements herein are provided only as background information in connection with this disclosure and do not necessarily constitute prior art.

[0003] Human SEZ6 (epilepsy-associated protein 6, hereinafter referred to as hSEZ6) is a single-transmembrane protein, mainly expressed in brain tissue, with almost no expression in the periphery. In the field of neurological diseases, knockout of SEZ6 in mice promotes the formation of short, multi-synapses (Neuron 56, 621–639, November 21, 2007); SEZ6 is a substrate of BACE1, and soluble SEZ6 in cerebrospinal fluid can serve as a marker for Alzheimer's disease (Molecular Neurodegeneration 2016 11:67).

[0004] SEZ6, as a tumor-specific marker, is specifically highly expressed in neuroendocrine tumor tissues. Immunohistochemical analysis of clinical specimens shows that SEZ6 is highly expressed in small cell lung cancer, and the expression level is much higher than in normal tissues. In tumor tissue sections from 174 small cell lung cancer patients, approximately 70% of the tumors showed positive expression of SEZ6 (published patent: US2016287720A1).

[0005] Antibody-drug conjugates (ADCs) link monoclonal antibodies or antibody fragments to biologically active cytotoxins via stable chemical linker compounds. This fully leverages the specificity of antibodies in binding to tumor cell surface antigens and the high efficiency of cytotoxins, while avoiding the drawbacks of the former's low efficacy and the latter's excessive toxicity. This means that, compared to traditional chemotherapy drugs, antibody-drug conjugates can precisely bind to tumor cells and reduce the impact on normal cells (Mullard A, (2013) Nature Reviews Drug Discovery, 12:329–332; DiJoseph JF, Armellino DC, (2004) Blood, 103:1807-1814).

[0006] Based on the aforementioned high specific expression of SEZ6 in tumors, AbbVie Pharmaceuticals developed an ADC targeting SEZ6, ABBV-011, for the indication of small cell lung cancer (public patent: WO2013126810A). It utilizes calic acid as a toxin, has a DAR value of 2, and employs a non-degradable linker. This ADC entered Phase I clinical trials in 2019 (NCT03639194). However, further development of more effective ADC drugs is still needed.

[0007] However, ADCs have a more complex heterogeneous structure than antibodies, which poses a greater challenge to ADC formulations for therapeutic purposes. Summary of the Invention

[0008] This disclosure provides a pharmaceutical composition comprising an antibody-drug conjugate and a buffer, said antibody-drug conjugate having the following structure:

[0009] in:

[0010] Pc is an anti-SEZ6 antibody or its antigen-binding fragment, comprising a heavy chain variable region and a light chain variable region, wherein: the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 03, SEQ ID NO: 04 and SEQ ID NO: 05 respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 07 and SEQ ID NO: 08 respectively;

[0011] n is an integer or decimal from 1 to 8;

[0012] The buffers are histidine buffers, tris(hydroxymethyl)aminomethane (Tris) buffers, phosphate buffers, acetate buffers, citrate buffers, succinate buffers, tartrate buffers, fumarate buffers, and glycylglycine buffers.

[0013] In an optional implementation, the buffer is a histidine buffer.

[0014] In an optional embodiment, the buffer is a histidine-histidine hydrochloride buffer, a histidine-citric acid buffer, or a histidine-hydrochloride buffer.

[0015] In some embodiments, the anti-SEZ6 antibody or its antigen-binding fragment includes a heavy chain variable region as shown in SEQ ID NO: 01 and a light chain variable region as shown in SEQ ID NO: 02.

[0016] In some embodiments, the anti-SEZ6 antibody or its antigen-binding fragment comprises a heavy chain as shown in SEQ ID NO: 09 and a light chain as shown in SEQ ID NO: 10.

[0017] In some implementations, n is an integer or decimal from 2 to 8; preferably, n is an integer or decimal from 4 to 8, 2 to 7, 2 to 6, 3 to 7, 3 to 6, or 4 to 7; more preferably, n is an integer or decimal from 4 to 6.

[0018] In an optional implementation, n is approximately 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0.

[0019] In some embodiments, the pH of the pharmaceutical composition is 4.5 to 6.5; preferably, the pH of the pharmaceutical composition is 4.8 to 6.2; more preferably, the pH of the pharmaceutical composition is 5.0 to 6.0.

[0020] In an optional embodiment, the pH of the pharmaceutical composition is 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, and any range between these values; preferably, the pH is about 5.5, where "about" means ±0.2, such as about 5.5 being 5.5 ± 0.2 (i.e., pH 5.3 to 5.7).

[0021] In some embodiments, the concentration of the buffer is from 5 mM to 50 mM, preferably from 10 mM to 30 mM, and more preferably about 20 mM. The "about" is ±10%, for example, a buffer concentration of about 20 mM is 20 mM ± 2 mM. Non-limiting embodiments include 5 mM, 10 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 30 mM, and any range between these values.

[0022] In some embodiments, the pharmaceutical composition comprises a histidine buffer at a concentration of 5 mM to 50 mM, preferably 10 mM to 30 mM, and more preferably about 20 mM. The "about" preferably refers to ±10%, such as about 20 mM of histidine buffer, which is equivalent to 20 mM ± 2 mM of histidine buffer.

[0023] In some embodiments, the pharmaceutical composition comprises a histidine-histidine hydrochloride buffer at a concentration of 5 mM to 50 mM, preferably 10 mM to 30 mM, and more preferably about 20 mM. The "about" preferably refers to ±10%, such as about 20 mM of histidine-histidine hydrochloride buffer, which is equivalent to 20 mM ± 2 mM of histidine buffer.

[0024] In some embodiments, the pharmaceutical composition comprises a histidine-histidine hydrochloride buffer at a concentration of about 10 mM. In some embodiments, the pharmaceutical composition comprises a histidine-histidine hydrochloride buffer at a concentration of about 20 mM. In some embodiments, the pharmaceutical composition comprises a histidine-histidine hydrochloride buffer at a concentration of about 30 mM. In some embodiments, the pharmaceutical composition comprises a histidine-histidine hydrochloride buffer at a concentration of 10 mM. In some embodiments, the pharmaceutical composition comprises a histidine-histidine hydrochloride buffer at a concentration of 20 mM. In some embodiments, the pharmaceutical composition comprises a histidine-histidine hydrochloride buffer at a concentration of 30 mM.

[0025] In some embodiments, the pharmaceutical composition further comprises a surfactant.

[0026] In an optional embodiment, the surfactant is selected from polysorbates (e.g., polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80), poloxamer, polyhydroxyanisole, Triton, sodium dodecyl sulfonate, sodium lauryl sulfonate, sodium octyl glycoside, lauryl-sulfobetaine, myristyl-sulfobetaine, linoleyl-sulfobetaine, stearyl-sulfobetaine, lauryl-sarcosine, myristyl-sarcosine, linoleyl-sarcosine, stearyl-sarcosine, linoleyl-betaine Bases, myristyl-betaine, cetyl-betaine, lauramide propyl-betaine, cocaramide propyl-betaine, linoleamide propyl-betaine, myristamide propyl-betaine, palmitoamide propyl-betaine, isostearamide propyl-betaine, myristamide propyl-dimethylamine, palmitoamide propyl-dimethylamine, isostearamide propyl-dimethylamine, sodium methyl cocoyl, sodium methyl oleyl taurate, polyethylene glycol, polypropylene glycol, and copolymers of ethylene and propylene glycol, etc.

[0027] In an optional embodiment, the surfactant is polysorbate 20, polysorbate 40, polysorbate 60, or polysorbate 80, preferably polysorbate 80.

[0028] In some embodiments, the surfactant concentration is from 0.01 mg / mL to 1.0 mg / mL, preferably from 0.1 mg / mL to 0.8 mg / mL, more preferably from 0.2 mg / mL to 0.6 mg / mL, and even more preferably about 0.4 mg / mL. The "about" preferably refers to ±10%, such as a surfactant concentration of about 0.4 mg / mL, i.e., 0.4 mg / mL ± 0.04 mg / mL. Non-limiting examples include 0.01 mg / mL, 0.02 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.15 mg / mL, 0.2 mg / mL, 0.25 mg / mL, 0.3 mg / mL, 0.35 mg / mL, 0.4 mg / mL, 0.45 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, 0.9 mg / mL, 1.0 mg / mL, and any range between these values.

[0029] In some embodiments, the surfactant concentration is about 0.4 mg / mL. In some embodiments, the surfactant concentration is about 0.6 mg / mL. In some embodiments, the surfactant concentration is about 0.2 mg / mL. In some embodiments, the surfactant concentration is 0.4 mg / mL. In some embodiments, the surfactant concentration is 0.6 mg / mL. In some embodiments, the surfactant concentration is 0.2 mg / mL.

[0030] In an optional embodiment, the pharmaceutical composition further comprises polysorbate 80 at a concentration of 0.01 mg / mL to 1.0 mg / mL, preferably 0.1 mg / mL to 0.8 mg / mL, more preferably 0.2 mg / mL to 0.6 mg / mL, and even more preferably about 0.4 mg / mL. The "about" is preferably ±10%, such as about 0.4 mg / mL of polysorbate 80, i.e., 0.4 mg / mL ± 0.04 mg / mL of polysorbate 80.

[0031] In some embodiments, the concentration of polysorbate 80 is about 0.4 mg / mL. In some embodiments, the concentration of polysorbate 80 is about 0.6 mg / mL. In some embodiments, the concentration of polysorbate 80 is about 0.2 mg / mL. In some embodiments, the concentration of polysorbate 80 is 0.4 mg / mL. In some embodiments, the concentration of polysorbate 80 is 0.6 mg / mL. In some embodiments, the concentration of polysorbate 80 is 0.2 mg / mL.

[0032] In some embodiments, the pharmaceutical composition further comprises a stabilizer.

[0033] In some embodiments, the stabilizer is selected from one or more of sugars, amino acids or their salts, and polyols. In some embodiments, the sugar is selected from one or more of sucrose, maltose, lactose, glucose, and trehalose. In some embodiments, the amino acid is selected from one or more of proline, aspartic acid, glutamic acid, lysine, arginine, glycine, and histidine. In some embodiments, the polyol is selected from one or more of mannitol and sorbitol.

[0034] In some embodiments, the stabilizer is selected from one or more of sucrose, trehalose, lactose, glucose, arginine or its salt, lysine or its salt, glycine or its salt, proline or its salt, mannitol, and sorbitol; more preferably, the stabilizer is selected from one or more of sucrose and arginine hydrochloride.

[0035] In some embodiments, the stabilizer is selected from sucrose.

[0036] In some embodiments, the stabilizer is selected from sucrose and arginine hydrochloride.

[0037] In some embodiments, the stabilizer concentration is from 10 mg / mL to 120 mg / mL, preferably from 20 mg / mL to 100 mg / mL, more preferably from 20 mg / mL to 80 mg / mL, and even more preferably from 20 mg / mL to 40 mg / mL or from 50 mg / mL to 80 mg / mL. Non-limiting examples include about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 7.2 mg / mL, about 7.4 mg / mL, about 7.6 mg / mL, about 7.8 mg / mL, about 8 mg / mL, about 8.2 mg / mL, about 8.4 mg / mL, about 8.6 mg / mL, about 8.8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 11 mg / mL, about 12 mg / mL, about 13 mg / mL, about 14 mg / mL, about 15 mg / mL, about 16 mg / mL, about 17 mg / mL, about 18 mg / mL, about 19 mg / mL, about 20 mg / mL, about 21 mg / mL, about 22 mg / mL, about 23 mg / mL, about 24 mg / mL, about 25 mg / mL. The values ​​are approximately 26 mg / mL, 27 mg / mL, 28 mg / mL, 29 mg / mL, 30 mg / mL, 31 mg / mL, 32 mg / mL, 33 mg / mL, 34 mg / mL, 35 mg / mL, 36 mg / mL, 37 mg / mL, 38 mg / mL, 39 mg / mL, 40 mg / mL, 41 mg / mL, 42 mg / mL, 43 mg / mL, 44 mg / mL, 45 mg / mL, 46 mg / mL, 47 mg / mL, 48 mg / mL, 49 mg / mL, 50 mg / mL, 55 mg / mL, 56 mg / mL, 60 mg / mL, 65 mg / mL, 70 mg / mL, 75 mg / mL, 80 mg / mL, or any value between any two. The optimal concentration is approximately 10%, such as approximately 75 mg / mL, which is 75 mg / mL ± 7.5 mg / mL.

[0038] In some embodiments, the pharmaceutical composition further comprises sucrose at a concentration of 10 mg / mL to 120 mg / mL. Preferably, it is sucrose at a concentration of 20 mg / mL to 100 mg / mL, more preferably 20 mg / mL to 80 mg / mL, and even more preferably 20 mg / mL to 40 mg / mL or 50 mg / mL to 80 mg / mL.

[0039] In some embodiments, the pharmaceutical composition further comprises arginine hydrochloride at a concentration of 10 mg / mL to 120 mg / mL. Preferably, it is arginine hydrochloride at a concentration of 20 mg / mL to 100 mg / mL, more preferably arginine hydrochloride at a concentration of 20 mg / mL to 80 mg / mL, and even more preferably arginine hydrochloride at a concentration of 20 mg / mL to 40 mg / mL.

[0040] In some embodiments, the pharmaceutical composition further comprises sucrose at a concentration of 10 mg / mL to 120 mg / mL and arginine hydrochloride at a concentration of 10 mg / mL to 120 mg / mL. Preferably, the concentration of sucrose is 20 mg / mL to 100 mg / mL and the concentration of arginine hydrochloride is 20 mg / mL to 100 mg / mL; more preferably, the concentration of sucrose is 20 mg / mL to 80 mg / mL and the concentration of arginine hydrochloride is 20 mg / mL to 80 mg / mL. Further preferably, the concentration of sucrose is 20 mg / mL to 40 mg / mL and the concentration of arginine hydrochloride is 20 mg / mL to 40 mg / mL.

[0041] In some embodiments, the pharmaceutical composition further comprises sucrose at a concentration of about 75 mg / mL. In some embodiments, the pharmaceutical composition further comprises sucrose at a concentration of 75 mg / mL. In some embodiments, the pharmaceutical composition further comprises sucrose at a concentration of about 60 mg / mL. In some embodiments, the pharmaceutical composition further comprises sucrose at a concentration of 60 mg / mL. In some embodiments, the pharmaceutical composition further comprises sucrose at a concentration of about 45 mg / mL. In some embodiments, the pharmaceutical composition further comprises sucrose at a concentration of 45 mg / mL.

[0042] In some embodiments, the pharmaceutical composition further comprises 20 mg / mL to 40 mg / mL of sucrose and 20 mg / mL to 40 mg / mL of arginine hydrochloride. In some embodiments, the pharmaceutical composition further comprises sucrose at a concentration of about 34.23 mg / mL and arginine hydrochloride at a concentration of about 21.07 mg / mL. In some embodiments, the pharmaceutical composition further comprises sucrose at a concentration of 34.23 mg / mL and arginine hydrochloride at a concentration of about 21 mg / mL. The "about" is preferably 10%, such as a sucrose concentration of about 34.23 mg / mL, which is 34.23 mg / mL ± 3.42 mg / mL.

[0043] In some embodiments, the pharmaceutical composition further comprises about 0.1 M sucrose and about 0.1 M arginine hydrochloride.

[0044] In some embodiments, the concentration of the antibody-drug conjugate in the pharmaceutical composition is from 10 mg / mL to 70 mg / mL, preferably from 10 mg / mL to 50 mg / mL, more preferably from 20 mg / mL to 50 mg / mL. Most preferably, it is about 20 mg / mL (18 mg / mL to 22 mg / mL). Specific, non-limiting examples include 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL, 15 mg / mL, 16 mg / mL, 17 mg / mL, 18 mg / mL, 19 mg / mL, 20 mg / mL, 21 mg / mL, 22 mg / mL, 23 mg / mL, 24 mg / mL, 25 mg / mL, 26 mg / mL, 27 mg / mL, 28 mg / mL, 29 mg / mL, 30 mg / mL, 35 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, and any range between these values. The optimal concentration is approximately 10%, such as approximately 20 mg / mL, which is 20 mg / mL ± 2 mg / mL.

[0045] In some embodiments, the concentration of the antibody-drug conjugate in the pharmaceutical composition is about 20 mg / mL.

[0046] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0047] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 5 mM to 50 mM, (c) a surfactant at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) a stabilizer at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0048] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0049] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 5 mM to 50 mM, (c) a surfactant at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) a stabilizer at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0050] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0051] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 10 mM to 30 mM, (c) a surfactant at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) a stabilizer at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0052] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0053] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 5 mM to 50 mM, (c) a surfactant at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) a stabilizer at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0054] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0055] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 5 mM to 50 mM, (c) a surfactant at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) a stabilizer at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0056] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0057] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histamine buffer at a concentration of 10 mM to 30 mM, (c) surfactant at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) stabilizer at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0058] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0059] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 10 mM to 30 mM, (c) a surfactant at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) a stabilizer at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0060] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0061] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 5 mM to 50 mM, (c) a surfactant at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) a stabilizer at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0062] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0063] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) a buffer at a concentration of 10 mM to 30 mM, (c) a surfactant at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) a stabilizer at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0064] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0065] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 5 mM to 50 mM, (c) polysorbate at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) sugar at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0066] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0067] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 10 mM to 30 mM, (c) polysorbate at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) sugar at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0068] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0069] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 5 mM to 50 mM, (c) polysorbate at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) sugar at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0070] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0071] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 5 mM to 50 mM, (c) polysorbate at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) sugar at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0072] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0073] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 10 mM to 30 mM, (c) polysorbate at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) sugar at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0074] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0075] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 10 mM to 30 mM, (c) polysorbate at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) sugar at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0076] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0077] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 5 mM to 50 mM, (c) polysorbate at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) sugar at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0078] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0079] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) histidine buffer at a concentration of 10 mM to 30 mM, (c) polysorbate at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) sugar at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0080] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0081] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) histidine buffer at a concentration of 10 mM to 30 mM, (c) polysorbate at a concentration of 0.1 mg / mL to 0.8 mg / mL, (d) sugar at a concentration of 20 mg / mL to 100 mg / mL, and (e) amino acid or salt thereof at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0082] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0083] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) histidine-histidine hydrochloride buffer at a concentration of 10 mM to 30 mM, (c) polysorbate 80 at a concentration of 0.1 mg / mL to 0.8 mg / mL, (d) sucrose at a concentration of 20 mg / mL to 100 mg / mL, and (e) arginine hydrochloride at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0084] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0085] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) histidine-histidine hydrochloride buffer at a concentration of 10 mM to 30 mM, (c) polysorbate 80 at a concentration of 0.2 mg / mL to 0.6 mg / mL, (d) sucrose at a concentration of 20 mg / mL to 40 mg / mL, and (e) arginine hydrochloride at a concentration of 20 mg / mL to 40 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0086] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0087] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) 20 mM histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL polysorbate 80, (d) 34.23 mg / mL sucrose, and (e) 21.07 mg / mL arginine hydrochloride, wherein the pH of the composition is from 4.5 to 6.5.

[0088] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0089] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) 20 mM histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL polysorbate 80, (d) 0.1 M sucrose, and (e) 0.1 M arginine hydrochloride, wherein the pH of the composition is from 4.5 to 6.5.

[0090] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0091] (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 10 mM to 30 mM of histidine-histidine hydrochloride buffer, (c) 0.2 mg / mL to 0.6 mg / mL of polysorbate 80, and (d) 20 mg / mL to 80 mg / mL of sucrose, wherein the pH of the composition is 4.5 to 6.5.

[0092] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0093] (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 10 mM to 30 mM of histidine-histidine hydrochloride buffer, (c) 0.2 mg / mL to 0.6 mg / mL of polysorbate 80, and (d) 50 mg / mL to 80 mg / mL of sucrose, wherein the pH of the composition is 4.5 to 6.5.

[0094] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0095] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) polysorbate 80 at a concentration of 0.2 mg / mL to 0.6 mg / mL, and (d) sucrose at a concentration of 20 mg / mL to 80 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0096] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0097] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) polysorbate 80 at a concentration of 0.2 mg / mL to 0.6 mg / mL, and (d) sucrose at a concentration of 50 mg / mL to 80 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0098] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0099] (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 10 mM to 30 mM of histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL of polysorbate 80, and (d) 20 mg / mL to 80 mg / mL of sucrose, wherein the pH of the composition is 4.5 to 6.5.

[0100] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0101] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) histidine-histidine hydrochloride buffer at a concentration of 10 mM to 30 mM, (c) polysorbate 80 at a concentration of about 0.4 mg / mL, and (d) sucrose at a concentration of 50 mg / mL to 80 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0102] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0103] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) histidine-histidine hydrochloride buffer at a concentration of 10 mM to 30 mM, (c) polysorbate 80 at a concentration of 0.2 mg / mL to 0.6 mg / mL, and (d) sucrose at a concentration of about 75 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0104] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0105] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL of polysorbate 80, and (d) 20 mg / mL to 80 mg / mL of sucrose, wherein the pH of the composition is 4.5 to 6.5.

[0106] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0107] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL of polysorbate 80, and (d) 50 mg / mL to 80 mg / mL of sucrose, wherein the pH of the composition is 4.5 to 6.5.

[0108] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0109] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) polysorbate 80 at a concentration of 0.2 mg / mL to 0.6 mg / mL, and (d) about 75 mg / mL of sucrose, wherein the pH of the composition is 4.5 to 6.5.

[0110] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0111] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) histidine-histidine hydrochloride buffer at a concentration of 10 mM to 30 mM, (c) polysorbate 80 at a concentration of about 0.4 mg / mL, and (d) sucrose at a concentration of about 75 mg / mL, wherein the pH of the composition is from 4.5 to 6.5.

[0112] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0113] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL of polysorbate 80, and (d) about 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0114] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0115] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL of polysorbate 80, and (d) about 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0116] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0117] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL of polysorbate 80, and (d) about 40 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0118] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0119] (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, (d) 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0120] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0121] (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, (d) 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0122] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0123] (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, (d) 40 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0124] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0125] (a) 20 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0126] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0127] (a) 30 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0128] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0129] (a) 40 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0130] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0131] (a) 50 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0132] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0133] (a) 60 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0134] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0135] (a) 20 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0136] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0137] (a) 30 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0138] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0139] (a) 40 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0140] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0141] (a) 50 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0142] In an optional embodiment, the aforementioned pharmaceutical composition comprises the following components:

[0143] (a) 60 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0144] This disclosure also provides a lyophilized formulation containing an antibody-drug conjugate, characterized in that the formulation, upon reconstitution, can form the pharmaceutical composition described above.

[0145] This disclosure also provides a lyophilized formulation containing an antibody-drug conjugate, characterized in that the lyophilized formulation is obtained by freeze-drying the aforementioned pharmaceutical composition.

[0146] This disclosure also provides a method for preparing a lyophilized formulation containing an antibody-drug conjugate, including the step of lyophilizing the pharmaceutical composition as described above.

[0147] In an optional embodiment, the freeze-drying process in the method for preparing a lyophilized formulation comprising an antibody-drug conjugate sequentially includes pre-freezing, primary drying, and secondary drying steps. Freeze-drying is performed by freezing the formulation and subsequently sublimating water at a temperature suitable for primary drying. Under these conditions, the product temperature is below the eutectic point or collapse temperature of the formulation. Typically, the primary drying temperature ranges from about -45 to 25°C (assuming the product remains frozen during the primary drying process). The size and type of the formulation, the container holding the sample (e.g., a glass vial), and the volume of liquid determine the required drying time, which can range from several hours to several days (e.g., 30-60 hours). The secondary drying stage can be carried out at about 0-40°C, depending primarily on the type and size of the container and the type of protein used. The secondary drying time is determined by the desired residual moisture level in the product and typically requires at least about 5 hours. Typically, the water content of the low-pressure freeze-dried formulation is less than about 5%, preferably less than about 3%.

[0148] In one optional embodiment of this disclosure, the freeze-drying process includes the following steps:

[0149] (1) Pre-freeze at -2℃ to -10℃ for 1 to 180 minutes;

[0150] (2) Pre-freeze at -15℃ to -45℃ for 1 to 180 minutes;

[0151] (3) Dry at -10℃ to -20℃ and 0.01-1mbar for 2100 to 2700 minutes;

[0152] (4) Dry at -10℃ to -20℃ and 0.01-1mbar for 0 to 60 minutes;

[0153] (5) Dry at 20℃-30℃ and 0.01-1mbar for 480-900 minutes.

[0154] In one optional embodiment of this disclosure, the freeze-drying process includes the following steps:

[0155] (1) Pre-freeze at -5℃ for 1 to 180 minutes;

[0156] (2) Pre-freeze at -40℃ for 1 to 180 minutes;

[0157] (3) Dry at -15℃ and 0.01-1mbar for 2100-2700 minutes;

[0158] (4) Dry at -15℃ and 0.01-1mbar for 0 to 60 minutes;

[0159] (5) Dry at 25℃ and 0.01-1mbar for 480-900 minutes.

[0160] In one optional embodiment of this disclosure, the freeze-drying process includes the following steps:

[0161] (1) Pre-freeze at -5℃ for 120 minutes;

[0162] (2) Pre-freeze at -40℃ for 120 minutes or 150 minutes;

[0163] (3) Dry at -15℃ and 0.01-1mbar for 1800 minutes, 2100 minutes, or 2580 minutes;

[0164] (4) Dry at -15℃ and 0.01-1mbar for 0 minutes;

[0165] (5) Dry at 25℃ and 0.01-1mbar for 360 and 660 minutes.

[0166] In some embodiments, the lyophilized formulation is stable at 2-8°C for at least 16 days, at least 1 month, at least 3 months, at least 6 months, at least 12 months, at least 18 months, or at least 24 months. In some embodiments, the lyophilized formulation is stable at 40°C for at least 7 days, at least 14 days, at least 28 days, or at least 30 days.

[0167] This disclosure also provides a reconstituted solution containing an antibody-drug conjugate, characterized in that the reconstituted solution is prepared by reconstituted a lyophilized formulation as described above.

[0168] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0169] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 5 mM to 50 mM, (c) a surfactant at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) a stabilizer at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0170] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0171] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 5 mM to 50 mM, (c) a surfactant at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) a stabilizer at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0172] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0173] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 10 mM to 30 mM, (c) a surfactant at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) a stabilizer at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0174] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0175] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 5 mM to 50 mM, (c) a surfactant at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) a stabilizer at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0176] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0177] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 5 mM to 50 mM, (c) a surfactant at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) a stabilizer at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0178] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0179] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histamine buffer at a concentration of 10 mM to 30 mM, (c) surfactant at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) stabilizer at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0180] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0181] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 10 mM to 30 mM, (c) a surfactant at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) a stabilizer at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0182] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0183] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 5 mM to 50 mM, (c) a surfactant at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) a stabilizer at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0184] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0185] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) a buffer at a concentration of 10 mM to 30 mM, (c) a surfactant at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) a stabilizer at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0186] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0187] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 5 mM to 50 mM, (c) polysorbate at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) sugar at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0188] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0189] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 10 mM to 30 mM, (c) polysorbate at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) sugar at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0190] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0191] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 5 mM to 50 mM, (c) polysorbate at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) sugar at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0192] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0193] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 5 mM to 50 mM, (c) polysorbate at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) sugar at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0194] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0195] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 10 mM to 30 mM, (c) polysorbate at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) sugar at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0196] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0197] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 10 mM to 30 mM, (c) polysorbate at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) sugar at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0198] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0199] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 5 mM to 50 mM, (c) polysorbate at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) sugar at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0200] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0201] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) histidine buffer at a concentration of 10 mM to 30 mM, (c) polysorbate at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) sugar at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0202] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0203] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) histidine buffer at a concentration of 10 mM to 30 mM, (c) polysorbate at a concentration of 0.1 mg / mL to 0.8 mg / mL, (d) sugar at a concentration of 20 mg / mL to 100 mg / mL, and (e) amino acid or salt thereof at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0204] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0205] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) histidine-histidine hydrochloride buffer at a concentration of 10 mM to 30 mM, (c) polysorbate 80 at a concentration of 0.1 mg / mL to 0.8 mg / mL, (d) sucrose at a concentration of 20 mg / mL to 100 mg / mL, and (e) arginine hydrochloride at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0206] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0207] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) histidine-histidine hydrochloride buffer at a concentration of 10 mM to 30 mM, (c) polysorbate 80 at a concentration of 0.2 mg / mL to 0.6 mg / mL, (d) sucrose at a concentration of 20 mg / mL to 40 mg / mL, and (e) arginine hydrochloride at a concentration of 20 mg / mL to 40 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0208] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0209] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) 20 mM histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL polysorbate 80, (d) 34.23 mg / mL sucrose, and (e) 21.07 mg / mL arginine hydrochloride, wherein the pH of the composition is from 4.5 to 6.5.

[0210] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0211] (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) 20 mM histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL polysorbate 80, (d) 0.1 M sucrose, and (e) 0.1 M arginine hydrochloride, wherein the pH of the composition is from 4.5 to 6.5.

[0212] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0213] (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 10 mM to 30 mM of histidine-histidine hydrochloride buffer, (c) 0.2 mg / mL to 0.6 mg / mL of polysorbate 80, and (d) 20 mg / mL to 80 mg / mL of sucrose, wherein the pH of the composition is 4.5 to 6.5.

[0214] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0215] (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 10 mM to 30 mM of histidine-histidine hydrochloride buffer, (c) 0.2 mg / mL to 0.6 mg / mL of polysorbate 80, and (d) 50 mg / mL to 80 mg / mL of sucrose, wherein the pH of the composition is 4.5 to 6.5.

[0216] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0217] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) polysorbate 80 at a concentration of 0.2 mg / mL to 0.6 mg / mL, and (d) sucrose at a concentration of 20 mg / mL to 80 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0218] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0219] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) polysorbate 80 at a concentration of 0.2 mg / mL to 0.6 mg / mL, and (d) sucrose at a concentration of 50 mg / mL to 80 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0220] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0221] (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 10 mM to 30 mM of histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL of polysorbate 80, and (d) 20 mg / mL to 80 mg / mL of sucrose, wherein the pH of the composition is 4.5 to 6.5.

[0222] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0223] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) histidine-histidine hydrochloride buffer at a concentration of 10 mM to 30 mM, (c) polysorbate 80 at a concentration of about 0.4 mg / mL, and (d) sucrose at a concentration of 50 mg / mL to 80 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0224] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0225] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) histidine-histidine hydrochloride buffer at a concentration of 10 mM to 30 mM, (c) polysorbate 80 at a concentration of 0.2 mg / mL to 0.6 mg / mL, and (d) sucrose at a concentration of about 75 mg / mL, wherein the pH of the composition is 4.5 to 6.5.

[0226] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0227] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL of polysorbate 80, and (d) 20 mg / mL to 80 mg / mL of sucrose, wherein the pH of the composition is 4.5 to 6.5.

[0228] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0229] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL of polysorbate 80, and (d) 50 mg / mL to 80 mg / mL of sucrose, wherein the pH of the composition is 4.5 to 6.5.

[0230] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0231] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) polysorbate 80 at a concentration of 0.2 mg / mL to 0.6 mg / mL, and (d) about 75 mg / mL of sucrose, wherein the pH of the composition is 4.5 to 6.5.

[0232] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0233] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) histidine-histidine hydrochloride buffer at a concentration of 10 mM to 30 mM, (c) polysorbate 80 at a concentration of about 0.4 mg / mL, and (d) sucrose at a concentration of about 75 mg / mL, wherein the pH of the composition is from 4.5 to 6.5.

[0234] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0235] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL of polysorbate 80, and (d) about 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0236] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0237] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL of polysorbate 80, and (d) about 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0238] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0239] (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL of polysorbate 80, and (d) about 40 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0240] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0241] (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, (d) 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0242] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0243] (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, (d) 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0244] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0245] (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, (d) 40 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0246] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0247] (a) 20 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0248] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0249] (a) 30 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0250] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0251] (a) 40 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0252] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0253] (a) 50 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0254] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0255] (a) 60 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0256] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0257] (a) 20 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0258] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0259] (a) 30 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0260] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0261] (a) 40 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0262] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0263] (a) 50 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0264] In an optional embodiment, the aforementioned reconstitution solution comprises the following components:

[0265] (a) 60 mg / mL of the antibody-drug conjugate, (b) 20 mM of histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 60 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.0.

[0266] This disclosure also provides a method for preparing the above-mentioned reconstituted solution, including a step of reconstituted the aforementioned lyophilized preparation, wherein the solution used for reconstitution is selected from, but is not limited to, water for injection, physiological saline or glucose solution.

[0267] In some embodiments, the pharmaceutical composition or reconstituted solution as described in any of the preceding embodiments is an intravenous infusion preparation, an intravenous injection preparation, a subcutaneous injection preparation, an intraperitoneal injection preparation, or an intramuscular injection preparation. In some embodiments, the pharmaceutical composition or reconstituted solution as described in any of the preceding embodiments is an intravenous infusion preparation.

[0268] In some embodiments, the pharmaceutical composition or reconstituted solution as described in any of the preceding embodiments is suitable for intravenous infusion, intravenous injection, subcutaneous injection, intraperitoneal injection or intramuscular injection; preferably, it is suitable for intravenous infusion.

[0269] In some embodiments, the pharmaceutical composition, reconstituted solution, or lyophilized preparation described in any of the preceding embodiments is used to prepare a drug for intravenous infusion, intravenous injection, subcutaneous injection, intraperitoneal injection, or intramuscular injection; preferably, it is used to prepare a drug for intravenous infusion.

[0270] This disclosure also provides a medicine box that includes a container containing the pharmaceutical composition, lyophilized formulation, or reconstituted solution as described above.

[0271] In some implementations, the container is a neutral borosilicate glass vial for injection.

[0272] This disclosure also provides the use of the aforementioned pharmaceutical compositions or lyophilized formulations or reconstituted solutions or kits in the preparation of medicaments for the treatment or prevention of diseases.

[0273] In an optional implementation, the disease is a tumor or cancer.

[0274] In an optional implementation, the tumor or cancer is a tumor or cancer associated with SEZ6 expression.

[0275] In an optional implementation, the tumor or cancer is selected from lung cancer, breast cancer, liver cancer, hepatobiliary cancer, pancreatic cancer, stomach cancer, gastrointestinal cancer, intestinal cancer, colon cancer, colorectal cancer, kidney cancer, clear cell renal cell carcinoma, ovarian cancer, endometrial cancer, cervical cancer, bladder cancer, prostate cancer, testicular cancer, skin cancer, melanoma, leukemia, lymphoma, and bone cancer.

[0276] This disclosure also provides a pharmaceutical composition for an anti-SEZ6 antibody, wherein the anti-SEZ6 antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:3, SEQ ID NO:4 and SEQ ID NO:5 respectively; the light chain variable region comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:6, SEQ ID NO:7 and SEQ ID NO:8 respectively; the concentration of the anti-SEZ6 antibody is from about 1 mg / mL to about 100 mg / mL, preferably from about 10 mg / mL to about 100 mg / mL, more preferably from about 10 mg / mL to about 60 mg / mL, further preferably from about 10 mg / mL, about 20 mg / mL, about 30 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, and even more preferably from about 20 mg / mL; the "about" is preferably 10%, such that the concentration is preferably about 20 mg / mL, which is 20 mg / mL ± 2 mg / mL. In an optional embodiment, the concentration of the aforementioned anti-SEZ6 antibody is 20 mg / mL. Non-limiting examples include 20 mg / mL, 25 mg / mL, 30 mg / mL, 35 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, 55 mg / mL, 60 mg / mL, 65 mg / mL, 70 mg / mL, 75 mg / mL, 80 mg / mL, 85 mg / mL, 90 mg / mL, 95 mg / mL, 100 mg / mL, and any range between these values.

[0277] In an optional embodiment, the aforementioned anti-SEZ6 antibody comprises a heavy chain variable region as shown in SEQ ID NO: 01 and a light chain variable region as shown in SEQ ID NO: 02.

[0278] In an optional embodiment, the aforementioned anti-SEZ6 antibody comprises a heavy chain as shown in SEQ ID NO: 09 and a light chain as shown in SEQ ID NO: 10.

[0279] In an optional embodiment, the aforementioned anti-SEZ6 antibody pharmaceutical composition further comprises a histidine buffer, an acetate buffer, and a phosphate buffer, preferably a histidine-histidine hydrochloride buffer, an acetate-sodium acetate buffer, a disodium hydrogen phosphate-sodium dihydrogen phosphate buffer, and more preferably an acetate-sodium acetate buffer.

[0280] In an optional embodiment, the concentration of the aforementioned buffer is from about 10 mM to about 50 mM, preferably from about 10 mM to about 30 mM, more preferably from about 10 mM, about 15 mM, about 20 mM, about 25 mM, or about 30 mM.

[0281] In an optional embodiment, the concentration of the aforementioned buffer is approximately 10 mM.

[0282] In an optional embodiment, the pH of the aforementioned anti-SEZ6 antibody pharmaceutical composition is about 4.5 to 6.5, preferably about 4.5 to 5.5, and more preferably about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, about 5.0, about 5.1, about 5.2, or about 5.3.

[0283] In an optional embodiment, the pH of the aforementioned pharmaceutical composition is about 4.9.

[0284] In an optional embodiment, the aforementioned anti-SEZ6 antibody pharmaceutical composition further comprises a stabilizer. In an optional embodiment, the stabilizer is selected from sugars, amino acids or their salts, or polyols.

[0285] In an optional embodiment, the stabilizer is selected from sucrose, maltose, lactose, glucose, or trehalose. In an optional embodiment, the stabilizer is selected from arginine or a salt thereof, lysine or a salt thereof, glycine or a salt thereof, proline or a salt thereof. In an optional embodiment, the stabilizer is selected from mannitol or sorbitol.

[0286] In an optional embodiment, the stabilizer is selected from sucrose.

[0287] In an optional embodiment, the concentration of the aforementioned stabilizer is from 20 mg / mL to 100 mg / mL, preferably from 20 mg / mL to 80 mg / mL, more preferably from 50 mg / mL to 80 mg / mL, and even more preferably from about 50 mg / mL, about 60 mg / mL, about 70 mg / mL, or about 80 mg / mL.

[0288] In an optional embodiment, the concentration of the aforementioned stabilizer is approximately 60 mg / mL.

[0289] In an optional embodiment, the pharmaceutical composition of the anti-SEZ6 antibody as described above comprises:

[0290] (a) an anti-SEZ6 antibody at a concentration of 10 mg / mL to 100 mg / mL, (b) an acetate buffer at a concentration of 10 mM to 30 mM, and (c) a sugar at a concentration of 20 mM to 80 mM; the pH of the pharmaceutical composition is about 4.5 to 6.5.

[0291] In an optional embodiment, the pharmaceutical composition of the anti-SEZ6 antibody as described above comprises:

[0292] (a) an anti-SEZ6 antibody at a concentration of 10 mg / mL to 60 mg / mL, (b) an acetate buffer at a concentration of 10 mM to 30 mM, and (c) a sugar at a concentration of 50 mM to 80 mM; the pH of the pharmaceutical composition is about 4.5 to 6.5.

[0293] In an optional embodiment, the pharmaceutical composition of the anti-SEZ6 antibody as described above comprises:

[0294] (a) anti-SEZ6 antibody at a concentration of 10 mg / mL to 60 mg / mL, (b) 10 mM to 30 mM acetate-sodium acetate buffer, (c) 50 mM to 80 mM sucrose; the pH of the pharmaceutical composition is about 4.5 to 6.5.

[0295] In an optional embodiment, the pharmaceutical composition of the anti-SEZ6 antibody as described above comprises:

[0296] (a) anti-SEZ6 antibody at a concentration of 10 mg / mL to 60 mg / mL, (b) 10 mM to 30 mM acetate-sodium acetate buffer, (c) 50 mM to 80 mM sucrose; the pH of the pharmaceutical composition is about 4.5 to 5.5.

[0297] In an optional embodiment, the pharmaceutical composition of the anti-SEZ6 antibody as described above comprises:

[0298] (a) anti-SEZ6 antibody at a concentration of 10 mg / mL to 60 mg / mL, (b) about 10 mM acetate-sodium acetate buffer, (c) about 60 mM sucrose; the pH of the pharmaceutical composition is about 4.5 to 5.5.

[0299] In an optional embodiment, the pharmaceutical composition of the anti-SEZ6 antibody as described above comprises:

[0300] (a) anti-SEZ6 antibody at a concentration of 10 mg / mL to 60 mg / mL, (b) about 10 mM acetate-sodium acetate buffer, (c) about 50 mM sucrose; the pH of the pharmaceutical composition is about 4.5 to 5.5.

[0301] In an optional embodiment, the pharmaceutical composition of the anti-SEZ6 antibody as described above comprises:

[0302] (a) anti-SEZ6 antibody at a concentration of 10 mg / mL to 60 mg / mL, (b) about 10 mM acetate-sodium acetate buffer, (c) about 70 mM sucrose; the pH of the pharmaceutical composition is about 4.5 to 5.5.

[0303] In an optional embodiment, the pharmaceutical composition of the anti-SEZ6 antibody as described above comprises:

[0304] (a) anti-SEZ6 antibody at a concentration of 10 mg / mL to 60 mg / mL, (b) about 20 mM acetate-sodium acetate buffer, (c) about 60 mM sucrose; the pH of the pharmaceutical composition is about 4.5 to 5.5.

[0305] In an optional embodiment, the pharmaceutical composition of the anti-SEZ6 antibody as described above comprises:

[0306] (a) anti-SEZ6 antibody at a concentration of 10 mg / mL to 60 mg / mL, (b) about 30 mM acetate-sodium acetate buffer, (c) about 60 mM sucrose; the pH of the pharmaceutical composition is about 4.5 to 5.5.

[0307] In an optional embodiment, the pharmaceutical composition of the anti-SEZ6 antibody as described above comprises:

[0308] (a) anti-SEZ6 antibody at a concentration of 10 mg / mL to 60 mg / mL, (b) about 20 mM acetate-sodium acetate buffer, (c) about 70 mM sucrose; the pH of the pharmaceutical composition is about 4.5 to 5.5.

[0309] In an optional embodiment, the pharmaceutical composition of the anti-SEZ6 antibody as described above comprises:

[0310] (a) an anti-SEZ6 antibody at a concentration of 10 mg / mL to 60 mg / mL, (b) about 30 mM of acetate-sodium acetate buffer, and (c) about 70 mM of sucrose; the pH of the pharmaceutical composition is about 4.5 to 5.5. In an optional embodiment, the pharmaceutical composition of the anti-SEZ6 antibody as described above comprises:

[0311] (a) anti-SEZ6 antibody at a concentration of 10 mg / mL to 60 mg / mL, (b) about 20 mM acetate-sodium acetate buffer, (c) about 50 mM sucrose; the pH of the pharmaceutical composition is about 4.5 to 5.5.

[0312] In an optional embodiment, the pharmaceutical composition of the anti-SEZ6 antibody as described above comprises:

[0313] (a) anti-SEZ6 antibody at a concentration of 10 mg / mL to 60 mg / mL, (b) about 30 mM acetate-sodium acetate buffer, (c) about 50 mM sucrose; the pH of the pharmaceutical composition is about 4.5 to 5.5.

[0314] When point values ​​are mentioned in this disclosure, it should be understood that these point values ​​include a range of error. This range of error is due to factors such as laboratory environment, personnel operation, instruments, methodologies, and measurement errors.

[0315] As is well known to those skilled in the art, one, some, or all of the features of the various embodiments described in this disclosure can be further combined to form other embodiments of this disclosure. The above embodiments of this disclosure and other embodiments obtained by combination are further described in detail below.

[0316] Invention Details

[0317] This disclosure provides a pharmaceutical composition that is more conducive to manufacturing and administration, and has stable performance. Undesirable instabilities may include any one or more of the following: aggregation, deamidation (e.g., Asn deamidation), oxidation (e.g., Met oxidation), isomerization (e.g., Asp isomerization), clipping / hydrolysis / fragmentation (e.g., hinge region fragmentation), succinimide formation, unpaired cysteine ​​residues, toxin dissociation, etc. Specifically, the pharmaceutical composition described in this disclosure comprises an antibody-drug conjugate.

[0318] the term

[0319] To facilitate understanding of this disclosure, certain technical and scientific terms are specifically defined below. Unless otherwise expressly defined herein, all other technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0320] Antibody drug conjugates (ADCs) are antibodies linked to biologically active cytotoxins or small molecule drugs with cell-killing activity via linker units.

[0321] "Drug loading," also known as drug-to-antibody ratio (DAR), refers to the average number of drugs conjugated to each antibody in an ADC. It can range from, for example, about 1 to about 10 drugs conjugated to each antibody, and in some embodiments, from about 1 to about 8 drugs conjugated to each antibody, preferably from the ranges of 2-8, 2-7, 2-6, 2-5, 2-4, 3-4, 3-5, 5-6, 5-7, 5-8, and 6-8. Exemplarily, the drug loading can be an average of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. The general formula of the ADC disclosed herein includes a set of antibodies conjugated to drugs within the aforementioned range. In embodiments of the disclosure, the drug loading can be expressed as n. The drug loading can be determined using conventional methods such as UV / visible spectroscopy, mass spectrometry, ELISA assays, and HPLC.

[0322] The loading of cytotoxic drugs can be controlled using the following non-limiting methods, including:

[0323] (1) Control the molar ratio of the ligation reagent and the monoclonal antibody.

[0324] (2) Control the reaction time and temperature.

[0325] (3) Choose different reaction reagents.

[0326] The three-letter and single-letter codes for amino acids used in this disclosure are as described in J. biol. chem, 243, p3558 (1968).

[0327] The term "antibody" as used in this disclosure is used in the broadest sense and encompasses a variety of antibody structures, including but not limited to full-length antibodies and antibody fragments (or antigen-binding fragments, or antigen-binding portions), provided they exhibit the desired antigen-binding activity. Typically, a natural, complete antibody is a tetrapeptide chain structure consisting of two identical heavy chains and two identical light chains linked by interchain disulfide bonds.

[0328] The engineered antibody or antigen-binding fragments disclosed herein can be prepared and purified using conventional methods. For example, cDNA sequences encoding heavy and light chains can be cloned and recombined into GS expression vectors. Recombinant immunoglobulin expression vectors can stably transfect CHO cells. As a more recommended prior art, mammalian expression systems lead to glycosylation of antibodies, particularly at the highly conserved N-terminal site in the Fc region. Positive clones are scaled up in serum-free medium in a bioreactor to produce antibodies. The culture medium secreting antibodies can be purified using conventional techniques, such as using an A or G Sepharose FF column with adjusted buffer. Non-specifically bound components are washed away. The bound antibodies are then eluted using a pH gradient, and antibody fragments are detected by SDS-PAGE and collected. The antibodies can be concentrated by filtration using conventional methods. Soluble mixtures and polymers can also be removed using conventional methods, such as molecular sieving or ion exchange. The resulting product should be immediately frozen, e.g., at -70°C, or lyophilized.

[0329] "Buffer" refers to a buffer that tolerates pH changes through the action of its acid-base conjugate components. Examples of buffers that maintain pH within an appropriate range include acetates, succinates, gluconates, histidines, oxalates, lactates, phosphates, citrates, tartrates, fumarates, glycylglycine, and other organic acid buffers.

[0330] "Histidine buffer" is a buffer containing histidine ions. Examples of histidine salt buffers include histidine-hydrochloride, histidine-acetate, histidine-phosphate, histidine-sulfate, etc., with histidine-acetate buffer being preferred. Histidine-acetate buffer is prepared by reacting histidine with acetic acid, and histidine-hydrochloride buffer is prepared by reacting histidine with hydrochloric acid.

[0331] "Citrate buffer" is a buffer that includes citrate ions. Examples of citrate buffers include sodium citrate, potassium citrate, calcium citrate, magnesium citrate, etc. A preferred citrate buffer is sodium citrate.

[0332] "Succinate buffer" is a buffer containing succinate ions. Examples of succinate buffers include sodium succinate, potassium succinate, and calcium succinate. A preferred succinate buffer is sodium succinate. Exemplarily, the sodium succinate can be prepared from succinic acid and sodium hydroxide, or from succinic acid and sodium succinate.

[0333] A "phosphate buffer" is a buffer that contains phosphate ions. Examples of phosphate buffers include disodium hydrogen phosphate-sodium dihydrogen phosphate, disodium hydrogen phosphate-potassium dihydrogen phosphate, and disodium hydrogen phosphate-citric acid. A preferred phosphate buffer is disodium hydrogen phosphate-sodium dihydrogen phosphate.

[0334] "Acetate buffer" is a buffer that includes acetate ions. Examples of acetate buffers include sodium acetate, histidine acetate, potassium acetate, calcium acetate, magnesium acetate, etc. Sodium acetate is a preferred acetate buffer.

[0335] Poloxamer is a block copolymer of ethylene oxide and propylene oxide. It is water-soluble and used as a surfactant in pharmaceutical formulations. Examples of poloxamer include poloxamer 188 (P188).

[0336] "Pharmaceutical composition" refers to a mixture containing one or more antibody-drug conjugates or their physiologically / pharmacologically acceptable salts or prodrugs described herein, along with other chemical components, such as physiologically / pharmacologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to maintain the stability of the antibody active ingredient, facilitate administration to the organism, and enhance the absorption of the active ingredient to exert its biological activity.

[0337] In this disclosure, "pharmaceutical composition" and "formulation" are not mutually exclusive.

[0338] Unless otherwise specified, the solvent in the solution form of the pharmaceutical compositions described in this disclosure is water.

[0339] "Lyophilized formulation" refers to a pharmaceutical composition or formulation obtained by a vacuum freeze-drying step after the liquid or solution form has been processed.

[0340] As used herein, the terms “about” and “approximately” mean a numerical value within the acceptable margin of error of a specific value determined by a person skilled in the art, the numerical value depending in part on how it is measured or determined (i.e., the limits of the measurement system). For example, in every practice in the art, “about” may mean within or above 1 standard deviation. Alternatively, “about” or “substantially includes” may mean a range of up to 20%. Furthermore, particularly for biological systems or processes, the term may mean up to an order of magnitude or up to five times the numerical value. Unless otherwise stated, when a specific value appears in this application and claims, the meaning of “about” or “substantially includes” should be assumed to be within the acceptable margin of error of that specific value.

[0341] The pharmaceutical compositions disclosed herein achieve a stable effect: the antibody-drug conjugates therein substantially retain their physical and / or chemical stability and / or biological activity after storage; preferably, the pharmaceutical compositions substantially retain their physical and chemical stability and their biological activity after storage. The storage period is generally selected based on the intended shelf life of the pharmaceutical composition. Currently, various analytical techniques are available for measuring protein stability, which can measure stability after storage at a selected temperature for a selected period of time.

[0342] A stable formulation is one in which no significant changes are observed when stored at refrigerated temperatures (2-8°C) for at least 3 months, preferably 6 months, more preferably 1 year, and even more preferably up to 2 years. Additionally, stable liquid formulations include those that exhibit the desired characteristics after storage at temperatures including 25°C for periods of 1 month, 3 months, and 6 months. Typical examples of stability include: aggregation or degradation of antibody monomers typically not exceeding about 10%, preferably not exceeding about 5%, as determined by SEC-HPLC. Visually, the formulation is a pale yellow, nearly colorless, clear liquid or colorless, or clear to slightly milky white. The concentration, pH, and osmotic pressure of the formulation exhibit variations not exceeding ±10%. A reduction of not more than about 10%, preferably not more than about 5%, is typically observed. Aggregation typically forms at a rate of not more than about 10%, preferably not more than about 5%.

[0343] If, after visual inspection of color and / or clarity, or by means of UV light scattering, size exclusion chromatography (SEC), and dynamic light scattering (DLS), the antibody-drug conjugate does not show significant increase in aggregation, precipitation, and / or denaturation, then the antibody-drug conjugate "retains its physical stability" in the pharmaceutical formulation. Changes in protein conformation can be evaluated by fluorescence spectroscopy (which determines the tertiary structure of the protein) and by FTIR spectroscopy (which determines the secondary structure of the protein).

[0344] If an antibody-drug conjugate does not exhibit significant chemical changes, then the antibody "retains its chemical stability" in the drug formulation. Chemical stability can be assessed by detecting and quantifying the chemically altered form of the protein. Degradation processes that frequently alter the chemical structure of proteins include hydrolysis or truncation (evaluated by methods such as size exclusion chromatography and CE-SDS), oxidation (evaluated by methods such as peptide mapping combined with mass spectrometry or MALDI / TOF / MS), deamidation (evaluated by methods such as ion exchange chromatography, capillary isoelectric focusing, peptide mapping, and isofpartate measurement), and isomerization (evaluated by measuring isofpartate content, peptide mapping, etc.).

[0345] If the biological activity of an antibody-drug conjugate at a given time is within a predetermined range of the biological activity exhibited when the drug formulation is prepared, then the antibody-drug conjugate "retains its biological activity" in the drug formulation.

[0346] "Optional" or "optionally" means that the event or circumstances described below may, but do not have to, occur, including the possibility that the event or circumstances may or may not occur. For example, "optionally contains 1-3 antibody heavy chain variable regions" means that the antibody heavy chain variable regions of a particular sequence may, but do not have to, be present.

[0347] For the preparation of conventional pharmaceutical compositions, please refer to the Chinese Pharmacopoeia.

[0348] The term "carrier" used in the context of this disclosure refers to a system that can alter the way a drug enters the body and its distribution within the body, control the rate of drug release, and deliver the drug to the target organ. Drug carrier release and targeting systems can reduce drug degradation and loss, decrease side effects, and improve bioavailability. For example, high-molecular-weight surfactants, due to their unique amphiphilic structure, can self-assemble to form various forms of aggregates, preferably such as micelles, microemulsions, gels, liquid crystals, and vesicles. These aggregates have the ability to encapsulate drug molecules while also exhibiting good membrane permeability, making them excellent drug carriers.

[0349] When applied to animals, humans, experimental subjects, cells, tissues, organs, or biological fluids, "giving" and "treatment" refer to the contact of an exogenous drug, therapeutic agent, diagnostic agent, or composition with the animal, human, subject, cell, tissue, organ, or biological fluid. "Giving" and "treatment" can refer to, for example, therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. Cellular treatment includes contact between a reagent and cells, as well as contact between a reagent and a fluid, wherein the fluid is in contact with the cells. "Giving" and "treatment" also mean the treatment of, for example, cells, by means of a reagent, diagnostic agent, conjugate composition, or by means of another cell in vitro and ex vivo. When applied to humans, veterinary, or research subjects, "treatment" refers to therapeutic treatment, preventative or prophylactic measures, research, and diagnostic applications.

[0350] "Treatment" means administering an oral or topical therapeutic agent, such as a composition comprising any of the compounds disclosed herein, to a patient who has symptoms of one or more diseases, and the therapeutic agent is known to have a therapeutic effect on these symptoms. Typically, a therapeutic agent is administered in a treated patient or population in an amount that effectively relieves symptoms of one or more diseases to induce the regression of such symptoms or inhibit their progression to any clinically measurable extent. The amount of a therapeutic agent that effectively relieves any specific disease symptom (also referred to as a "therapeuticly effective amount") can vary depending on a variety of factors, such as the patient's disease state, age, and weight, and the drug's ability to produce the desired therapeutic effect in the patient. Whether the disease symptoms have been relieved can be evaluated using any clinical test that a physician or other healthcare professional typically uses to assess the severity or progression of the symptoms. Although the embodiments disclosed herein (e.g., treatment methods or products) may be ineffective in alleviating symptoms of each target disease, they should reduce symptoms of the target disease in a statistically significant number of patients, as determined by any statistical test known in the art, such as the Student t-test, chi-square test, U-test according to Mann and Whitney, Kruskal-Wallis test (H-test), Jonckheere-Terpstra test, and Wilcoxon test.

[0351] An "effective amount" includes the amount sufficient to improve or prevent the symptoms or condition of a medically diagnosed disease. An effective amount also means the amount sufficient to allow or facilitate diagnosis. The effective amount for a particular patient or veterinary subject can vary depending on factors such as the condition to be treated, the patient's overall health, the route and dosage of administration, and the severity of side effects. An effective amount can be the maximum dose or administration regimen that avoids significant side effects or toxicity.

[0352] "Displacement" refers to the replacement of the solvent system in which the antibody protein is dissolved. For example, using a buffer system of a stabilizing formulation, a high-salt or hypertonic solvent system containing the antibody protein is replaced by a physical manipulation, thereby ensuring the antibody protein remains in the stabilizing formulation. The physical manipulation methods include, but are not limited to, ultrafiltration, dialysis, or reconstitution after centrifugation.

[0353] The reconstituted solutions disclosed herein may be administered by any suitable means, including parenteral, intrapulmonary, and intranasal administration, and, if local treatment is required, intralesional administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. Administration may be carried out via any suitable route, such as by injection, such as intravenous or subcutaneous injection. Various dosing schedules are considered herein, including, but not limited to, single or multiple administrations at multiple time points, bolus administration, and pulsatile infusion. In some embodiments, the reconstituted solutions disclosed herein are administered by subcutaneous injection. Attached Figure Description

[0354] Figure 1. Results of insoluble microparticles in formulation buffer system and pH screening.

[0355] Figure 2. Results of screening for insoluble microparticles as formulation stabilizers.

[0356] Figure 3. Results of insoluble microparticles in the lyophilization evaluation test of the formulation. Detailed Implementation

[0357] The following description, in conjunction with examples, further illustrates this disclosure, but these examples are not intended to limit the scope of this disclosure. Experimental methods in the examples of this disclosure that do not specify specific conditions are generally performed under conventional conditions, such as those described in Cold Spring Harbor Laboratory's *Antibody Technology Laboratory Manual* and *Molecular Cloning Handbook*; or under conditions recommended by the raw material or commercial manufacturer. Reagents whose specific source is not specified are commercially available, conventional reagents.

[0358] Preparation Example 1. Antibody Preparation

[0359] The SEZ6 antibody in this case is derived from antibody PR3178 of PCT international patent application WO2024230742A1 (included herein by reference in its entirety), and the relevant sequence is as follows:

[0360] 1) The CDR sequence of the anti-SEZ6 antibody is shown in the table below:

[0361] Table 1. CDR sequences obtained by the Kabat numbering rules

[0362] 2) Variable region sequence of humanized anti-SEZ6 antibody PR3178:

[0363] Anti-SEZ6 antibody heavy chain variable region:

[0364] Anti-SEZ6 antibody light chain variable region:

[0365] 3) The full-length sequence of the humanized anti-SEZ6 antibody PR3178 is as follows:

[0366] Anti-SEZ6 antibody heavy chain:

[0367] Anti-SEZ6 antibody light chain:

[0368] Preparation Example 2. Preparation of ADC

[0369] The pharmaceutical portion of the conjugate disclosed herein can be any suitable pharmaceutical product. Particularly suitable pharmaceutical products are described, for example, in PCT International Patent Application WO2024230742A1 (included herein by reference in its entirety).

[0370] The antibody-drug conjugate disclosed herein has the following structure:

[0371] Where n is 1 to 8, preferably 4 to 6.

[0372] The loading of cytotoxic drugs (n) can be controlled by non-restrictive methods, such as controlling the molar ratio of the drug linker fragment to the monoclonal antibody, controlling the reaction time and temperature, and selecting different reaction reagents.

[0373] The preferred embodiment n (DAR value) of the formulation in this disclosure is approximately 5.5, equivalent to 5.5 ± 0.5. The preferred embodiment n (DAR value) of the formulation in this disclosure is 5.5.

[0374] Example 1. Buffer system and pH screening of humanized anti-SEZ6 antibody

[0375] Based on the buffering range of common buffer salts and the isoelectric point of the humanized anti-SEZ6 antibody, two buffer systems were selected: 10 mM acetate-sodium acetate and histidine-histidine hydrochloride. Six systems with different pH values ​​and buffer salts were designed, with a pH range of 4.5–6.5. 6% (w / v) sucrose was used as the stabilizer, and the protein concentration was 20 mg / ml for formulation stability studies. Specific protocols are shown in Table 1.

[0376] Table 1. Buffer systems and pH screening schemes

[0377] I. Appearance

[0378] The stability of different formulations under high temperature (40℃), repeated freeze-thaw cycles (-35℃ / RT), and light exposure (4500-5500 lx) conditions was examined by testing the appearance of the samples.

[0379] The accelerated stability test results of DP01-DP06 at 40℃ are shown in Table 2. The results show that DP03 and DP06 exhibit slightly heavier opalescence at 0℃, indicating that these two systems are unfavorable for the appearance of the humanized anti-SEZ6 antibody. Comparison of acetate and histidine systems revealed that formulations DP01, DP02, DP04, and DP05 showed better appearance after 2 weeks at high temperature (40℃), indicating that the humanized anti-SEZ6 antibody has good stability in low-pH acetate systems (pH 4.5 and pH 5.0) and low-pH histidine systems (pH 5.0 and pH 5.5).

[0380] Table 2. Results of Formulation Development

[0381] II. Particle Size

[0382] The particle size of the samples was determined using high-throughput dynamic light scattering (DLS) to examine the particle size distribution and stability of different formulation systems. Larger particle sizes indicate that the proteins may aggregate or have poorer colloidal stability.

[0383] A 96-well plate was used. The appropriate amount of sample was added to the well using a pipette, and the presence of air bubbles was observed. If air bubbles were present, the plate was centrifuged. The particle size and the initial denaturation temperature (Tonset) were determined. The Tonset test required sealing with a sealing membrane. The particle size was measured at 25°C, and the sample volume was 100 ml. The results are shown in Table 3.

[0384] Table 3. Particle size detection results in formulation development

[0385] As shown in Table 3, the particle size of the DP01-06 prescription protein increases with increasing pH under all conditions, indicating that the anti-SEZ6 antibody may have poor colloidal stability in high pH systems.

[0386] III. Insoluble microparticles

[0387] The changes in insoluble microparticles in samples under repeated freeze-thaw conditions were detected using flow cytometry (MFI). A 96-well plate was used. A baseline test was first performed using purified water, followed by a preliminary optical adjustment test with 1 mL of sample. Only after passing this initial test were the samples analyzed. Data analysis was performed using filters to remove air bubbles and silicone oil. Finally, the concentrations of particles larger than 2 μm were statistically analyzed. The results are shown in Tables 4-5.

[0388] Table 4. Results of insoluble particulate matter detection in formulation development

[0389] Table 5. Results of insoluble particulate matter detection in formulation development

[0390] The results show that after repeated freeze-thaw cycles, the number of insoluble microparticles increased in all formulations. Among them, the increase in insoluble microparticles was more significant in DP05-06, i.e., the 10mM histidine buffer system, pH 6.0 and 6.5, and the 6% sucrose formulation. The number of insoluble microparticles with a particle size greater than 25μm in the DP06 formulation also increased significantly, while the increase in other formulations was smaller.

[0391] IV. Purity of SEC, iCIEF and NR-CE

[0392] Based on the molecular sieving mechanism of gel permeation chromatography columns, size exclusion chromatography (SEC) was used to separate monomers and polymers according to the molecular size of the analytes. At a detection wavelength of 280 nm, the proportions of impurity fragments and polymers were calculated using the area normalization method.

[0393] Table 6. Results of SEC-UPLC Detection of Monomers and Fragments in Formulation Development

[0394] Table 7. SEC-UPLC polymer assay results for formulation development

[0395] Table 6-7 summarizes the SEC purity results under different pH / buffer conditions. The results show that the DP04-06 formulation has a slightly higher polymer content than the DP01-03 formulation under all conditions, indicating that the anti-SEZ6 antibody has slightly lower SEC purity in the histidine system. Furthermore, after 2 weeks of high-temperature storage and 4 weeks of storage at 25°C, the proportion of fragments in the SEC purity of all formulations increased. Compared to other formulations, DP04 showed the most significant increase in fragments, while DP06 had a higher proportion of polymers.

[0396] The content of charge variants was determined using the iCIEF analytical method. A capillary was used as the separation channel. A DC voltage was applied across the capillary, creating a pH gradient within the amphoteric electrolyte solution. Each component migrated to its isoelectric point based on its charge difference, focusing into a very narrow segment, thus achieving component separation.

[0397] Table 8. iCIEF test results for prescription development Note: NT indicates that DP04 was detected (DP04's main peak was significantly lower than other groups at 0, and its fragments were significantly higher than other groups, so DP04 was screened out and not further detected).

[0398] Table 8 summarizes the iCIEF results under different pH buffer conditions. The results show that the acid peak content increased in all groups of samples after incubation at 40℃ for 2 weeks. There were no significant differences between different systems.

[0399] The purity of non-reduced CE-SDS was determined by capillary gel electrophoresis. Using a capillary as the separation channel and a high-voltage DC electric field as the driving force, separation was achieved based on the differences in the mobility (migration velocity per unit electric field strength) and / or partitioning behavior of each component in the sample.

[0400] Table 9. NRCE test results for prescription development

[0401] Table 9 summarizes the NRCE results under different pH / buffer conditions. The results show that at pH 0, the NRCE peak of DP03-04 was lower, with DP04 significantly lower than the other groups. After incubation at 40℃ for 2 weeks, the fragment content increased in all groups. Compared to other groups, the NRCE peak of DP03-04 decreased faster, and fragment growth was significant.

[0402] In summary, the anti-SEZ6 antibody generally performed well in terms of appearance, insoluble particles, and purity in 10mM acetate-sodium acetate systems at pH 4.5 and 5.0, and 6% sucrose. Since the performance was slightly worse at 10mM acetate-sodium acetate at pH 5.5, 10mM acetate-sodium acetate, pH 4.9 (pH 4.9 ± 0.4), 6% (w / v) sucrose, and a protein concentration of 20 mg / mL were selected as the preferred formulation.

[0403] Example 2. Buffer system and pH screening for anti-SEZ6 antibody-drug conjugate formulation

[0404] The design of buffer systems (histidine-histidine hydrochloride, acetate-sodium acetate, histidine-citric acid), pH (5.0, 5.5), and stabilizer types (sucrose, arginine hydrochloride) were used to screen the anti-SEZ6 antibody drug conjugate in different formulations under forced degradation conditions to determine its suitability for subsequent formulation and process research.

[0405] Sample batch number: 20231123. The sample was aseptically filled into 2mL vials, with a volume of 1.0mL / vial. The specific test protocol is shown in Table 10.

[0406] Table 10. Antibody-drug conjugate formulation buffer systems and pH screening protocols

[0407] The results of the formulation screening test for anti-SEZ6 antibody conjugates are shown in Tables 11-12 and Figure 1. The stability of the formulations was investigated under high temperature (40℃) and repeated freeze-thaw cycles to confirm the effects of pH, polysorbate 80 (Tween 80) content, buffer system, and ionic strength on the stability of the formulation. The formulation with the smallest trend of change was selected for subsequent process studies.

[0408] Table 11. Results of Formulation Buffer Systems and pH Screening for Anti-SEZ6 Antibody Drug Conjugates Note: " / " indicates not applicable. "TND" indicates not detected. "C" indicates colorless. "SO" indicates slightly opalescent liquid. "FP" indicates free of visible particles.

[0409] Table 12. Results of Formulation Buffer Systems and pH Screening for Anti-SEZ6 Antibody Drug Conjugates

[0410] The formulation screening test results showed that groups F3 and F4 exhibited a more pronounced increase in free toxins under accelerated high-temperature conditions. Among groups F1, F2, F5, F6, and F7, under accelerated high-temperature conditions, the percentage decrease in SEC peak purity, ranked from highest to lowest, was F7>F2>F5>F6>F1; the percentage increase in polymeric components, ranked from highest to lowest, was F7>F5>F6>F1>F2; and the percentage decrease in iCIEF peak purity, ranked from highest to lowest, was F5>F6>F7>F2>F1. Under repeated freeze-thaw conditions, group F7 showed a relatively significant increase in insoluble particulate matter. No significant differences were observed in appearance, DAR values, or other test results among the groups.

[0411] Based on the above data, group F1 (20 mmol / L histidine-histidine hydrochloride, pH 5.5, 0.04% (w / v) polysorbate 80, 7.5% (w / v) sucrose, and protein concentration 20 mg / ml) showed the smallest overall trend of change in each test item compared to other groups. Therefore, group F1 was selected as the basic formulation for subsequent formulation screening studies.

[0412] Example 3. Stabilizer Screening for Anti-SEZ6 Antibody Drug Conjugate Formulation

[0413] Based on the screening test results in Example 2, using 20 mmol / L histidine-histidine hydrochloride, pH 5.5, 0.04% (w / v) polysorbate 80, and 7.5% (w / ) sucrose as the base formulation, further screening schemes for stabilizers with different concentrations were designed. The stability of the antibody-drug conjugate formulation was investigated under accelerated conditions to determine the formulation of the antibody-drug conjugate formulation.

[0414] Sample batch number: 35240306F0101-101. The sample was aseptically filled into 2mL vials, with a volume of 1.0mL / vial. The specific test protocol is shown in Table 13.

[0415] Table 13. Screening scheme for antibody-drug conjugate formulation stabilizers

[0416] The results of the stabilizer screening test for the anti-SEZ6 antibody conjugate formulation are shown in Table 14 and Figure 2. The trends of key quality parameters among groups with different concentrations of sucrose were examined under accelerated conditions at 40℃. The optimal formulation was selected for subsequent process research.

[0417] Table 14. Screening results of stabilizers for formulations of anti-SEZ6 antibody-drug conjugates Note: " / " indicates not applicable. "TND" indicates not detected. "C" indicates colorless. "SO" indicates slightly opalescent liquid. "FP" indicates free of visible particles.

[0418] The formulation screening test results showed that, under high temperature conditions of 40℃, there were no significant differences in appearance among the groups. After 4 weeks of storage, group F3 showed the best performance in reduced CE purity and SEC purity. The decrease in the main component of reduced CE, ranked from highest to lowest, was: F2>F1>F3. The decrease in the monomer of SEC, ranked from highest to lowest, was: F1>F2>F3.

[0419] Taking all factors into consideration, group F3 (20 mmol / L histidine-histidine hydrochloride, pH 5.5, 7.5% (w / v) sucrose, 0.04% (w / v) polysorbate 80) was selected as the preferred formulation for subsequent process studies.

[0420] Example 4. Study on the freeze-drying process of anti-SEZ6 antibody-drug conjugate formulation

[0421] Based on the screening test results in Example 2, a further screening scheme for stabilizers with different concentrations was designed using 20 mmol / L histidine-histidine hydrochloride, pH 5.5, 0.04% (w / v) polysorbate 80, and 7.5% (w / v) sucrose as the base formulation. After lyophilization, stability was investigated to determine the antibody-drug conjugate formulation that can be used for lyophilization.

[0422] Sample batch number: 35240306F0101-101. The sample was aseptically filled into 2mL vials, with a volume of 1.0mL / vial. The specific experimental protocol is shown in Table 15, and the lyophilization parameters are shown in Table 16.

[0423] Table 15. Antibody-Drug Conjugate Formulation Lyophilization Evaluation Test Protocol

[0424] Table 16. Freeze-drying process parameters

[0425] The results of the lyophilization feasibility assessment of the anti-SEZ6 antibody-drug conjugate formulation are shown in Table 17 and Figure 3. The trends of key quality parameters among different groups were examined under accelerated conditions at 40℃ for lyophilized products with different concentrations of sucrose. The optimal formulation was selected for subsequent process research.

[0426] Table 17. Results of the Lyophilization Evaluation Test for Antibody-Drug Conjugate Formulations Note: " / " indicates not applicable. "TND" indicates not detected.

[0427] The results showed that there were significant differences in the changes of SEC purity and iCIEF purity among different groups of lyophilized products under accelerated conditions at 40℃, with group F3 showing the smallest change. No significant changes were observed among the groups for free toxins and insoluble particles.

[0428] Based on the results of the feasibility assessment test of the antibody-drug conjugate stock solution and lyophilization, the final formulation was confirmed as follows: 20.0 mg / mL antibody-drug conjugate, 20 mmol / L histidine-histidine hydrochloride (i.e., 0.72 mg / mL histidine and 3.22 mg / mL histidine hydrochloride monohydrate), 7.5% (w / v) sucrose, 0.04% (w / v) polysorbate 80, pH 5.5.

[0429] Example 5. Formulation confirmation of anti-SEZ6 antibody-drug conjugate formulation

[0430] Based on the results of the formulation screening and lyophilization feasibility assessment of the anti-SEZ6 antibody-drug conjugate, the preferred formulation was determined using 20 mmol / L histidine-histidine hydrochloride, pH 5.5, 7.5% (w / v) sucrose, and 0.04% (w / v) polysorbate 80.

[0431] The ultrafiltration recovery sample of the anti-SEZ6 antibody-drug conjugate (batch number: 35240409F0101) was selected for stock solution preparation. Lyophilization was performed according to the proposed lyophilization process parameters in Table 18. The stability of the antibody-drug conjugate stock solution and lyophilized product was investigated under accelerated and long-term conditions, respectively. The stability protocols are detailed in Table 19. A summary of the formulation confirmation data is shown in Tables 20-21.

[0432] Table 18. Freeze-drying process parameters

[0433] Table 19. Antibody-Drug Conjugate Formulation Validation Test Protocol

[0434] Table 20. Results of Formulation Validation Tests for Antibody-Drug Conjugates Note: " / " indicates not applicable. "C" indicates colorless. "SO" indicates slightly opalescent liquid. "FP" indicates free of visible particles.

[0435] Table 21. Results of Formulation Validation Tests for Antibody-Drug Conjugates

[0436] The experimental results showed that the quality changes of the antibody-drug conjugate stock solution and the lyophilized product of the optimized formulation (20 mmol / L histidine-histidine hydrochloride, pH 5.5, 7.5% (w / v) sucrose, 0.04% (w / v) polysorbate 80) under accelerated conditions at 25℃ and 40℃ were not significantly different from those observed in the previous screening and lyophilization feasibility assessment. This optimized formulation exhibited good stability in both the stock solution and the lyophilized product, and can be used for the preparation of anti-SEZ6 antibody-drug conjugates.

[0437] Although specific embodiments of the invention have been described in detail, those skilled in the art will understand that various modifications and variations can be made to the details based on all the published teachings, and all such changes are within the scope of protection of the invention. The entire scope of the invention is given by the appended claims and any equivalents thereof.

Claims

1. A pharmaceutical composition comprising an antibody drug conjugate and a buffer, the antibody drug conjugate having the structure shown below: in: Pc is an anti-SEZ6 antibody or its antigen-binding fragment, comprising a heavy chain variable region and a light chain variable region, wherein: the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 03, SEQ ID NO: 04 and SEQ ID NO: 05 respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 07 and SEQ ID NO: 08 respectively; n is an integer or decimal from 1 to 8; The buffer is a histidine buffer, a tris(hydroxymethyl)aminomethane (Tris) buffer, a phosphate buffer, an acetate buffer, a citrate buffer, or a succinate buffer. Preferably, the buffer is a histidine-hydrochloride histidine buffer, a histidine-citric acid buffer, or a histidine-hydrochloride buffer.

2. The pharmaceutical composition of claim 1, wherein, The anti-SEZ6 antibody or its antigen-binding fragment includes a heavy chain variable region as shown in SEQ ID NO: 01 and a light chain variable region as shown in SEQ ID NO: 02; Preferably, the anti-SEZ6 antibody or its antigen-binding fragment comprises a heavy chain as shown in SEQ ID NO: 09 and a light chain as shown in SEQ ID NO:

10.

3. The pharmaceutical composition according to claim 1 or 2, characterized in that, The n is an integer or decimal from 2 to 8; preferably, n is an integer or decimal from 4 to 8; more preferably, n is an integer or decimal from 4 to 6; even more preferably, n is 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, or 6.

0.

4. The pharmaceutical composition according to any one of claims 1 to 3, characterized in that, The pH of the pharmaceutical composition is 4.5 to 6.5; preferably, the pH of the pharmaceutical composition is 4.8 to 6.2; more preferably, the pH of the pharmaceutical composition is 5.0 to 6.

0.

5. The pharmaceutical composition according to any one of claims 1-4, characterized in that, The concentration of the buffer is 5 mM to 50 mM, preferably 10 mM to 30 mM, and more preferably about 20 mM.

6. The pharmaceutical composition according to any one of claims 1 to 5, characterized in that, The pharmaceutical composition further comprises a surfactant, preferably polysorbate or poloxamer, more preferably polysorbate 80, polysorbate 20, polysorbate 40, or polysorbate 60, and even more preferably polysorbate 80.

7. The pharmaceutical composition of claim 6, wherein, The surfactant concentration is from 0.01 mg / mL to 1.0 mg / mL, preferably from 0.1 mg / mL to 0.8 mg / mL, more preferably from 0.2 mg / mL to 0.6 mg / mL, and even more preferably about 0.4 mg / mL.

8. The pharmaceutical composition according to any one of claims 1 to 7, characterized in that, The pharmaceutical composition further comprises a stabilizer, preferably selected from one or more of sugars, amino acids or their salts, and polyols; more preferably, the stabilizer is selected from one or more of sucrose, trehalose, lactose, glucose, arginine or its salts, lysine or its salts, glycine or its salts, proline or its salts, mannitol, and sorbitol; even more preferably, the stabilizer is selected from one or more of sucrose and arginine hydrochloride.

9. The pharmaceutical composition of claim 8, wherein, The stabilizer concentration is from 10 mg / mL to 120 mg / mL, preferably from 20 mg / mL to 100 mg / mL, more preferably from 20 mg / mL to 80 mg / mL, and even more preferably from about 20 mg / mL, about 21 mg / mL, about 22 mg / mL, about 33 mg / mL, about 34 mg / mL, about 35 mg / mL, about 54 mg / mL, about 55 mg / mL, about 56 mg / mL, or about 75 mg / mL.

10. The pharmaceutical composition according to any one of claims 1 to 9, characterized in that, The concentration of the antibody-drug conjugate is from 10 mg / mL to 70 mg / mL, preferably from 10 mg / mL to 50 mg / mL, more preferably from 20 mg / mL to 50 mg / mL, and even more preferably from about 20 mg / mL, about 30 mg / mL, about 40 mg / mL or about 50 mg / mL.

11. The pharmaceutical composition according to any one of claims 1 to 10, characterized in that, It contains the following components: (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 5 mM to 50 mM, (c) a surfactant at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) a stabilizer at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.

5. Preferably, the pharmaceutical composition comprises the following components: (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 5 mM to 50 mM, (c) polysorbate at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) sugar at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.

5. Preferably, the pharmaceutical composition comprises the following components: (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) histidine buffer at a concentration of 10 mM to 30 mM, (c) polysorbate at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) sugar at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the composition is 4.5 to 6.

5. More preferably, the pharmaceutical composition comprises the following components: (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 10 mM to 30 mM of histidine-histidine hydrochloride buffer, (c) 0.2 mg / mL to 0.6 mg / mL of polysorbate 80, (d) 20 mg / mL to 80 mg / mL of sucrose, wherein the pH of the composition is 4.5 to 6.5; More preferably, the pharmaceutical composition comprises the following components: (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM of histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL of polysorbate 80, and (d) about 75 mg / mL of sucrose, wherein the pH of the composition is 5.0 to 6.

0.

12. A lyophilized formulation comprising an antibody drug conjugate, characterized in that, The lyophilized formulation, upon reconstitution, can form the pharmaceutical composition according to any one of claims 1 to 11.

13. A lyophilized formulation comprising an antibody drug conjugate, characterized in that, The lyophilized formulation is obtained by freeze-drying the pharmaceutical composition according to any one of claims 1 to 11.

14. A method of preparing a lyophilized formulation containing an antibody drug conjugate, characterized in that, The step includes freeze-drying the pharmaceutical composition according to any one of claims 1 to 11.

15. A reconstituted solution comprising an antibody drug conjugate, wherein the reconstituted solution is substantially free of a residual solvent. The reconstituted solution is prepared by reconstituted the lyophilized formulation according to claim 12 or 13, and the reconstituted solution comprises the following components: (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) a buffer at a concentration of 5 mM to 50 mM, (c) a surfactant at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) a stabilizer at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.

5. Preferably, the reconstituted solution comprises the following components: (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 70 mg / mL, (b) histidine buffer at a concentration of 5 mM to 50 mM, (c) polysorbate at a concentration of 0.01 mg / mL to 1.0 mg / mL, and (d) sugar at a concentration of 10 mg / mL to 120 mg / mL, wherein the pH of the composition is 4.5 to 6.

5. Preferably, the reconstituted solution comprises the following components: (a) the antibody-drug conjugate at a concentration of 10 mg / mL to 50 mg / mL, (b) histidine buffer at a concentration of 10 mM to 30 mM, (c) polysorbate at a concentration of 0.1 mg / mL to 0.8 mg / mL, and (d) sugar at a concentration of 20 mg / mL to 100 mg / mL, wherein the pH of the reconstituted solution is 4.5 to 6.5; More preferably, the reconstituted solution comprises the following components: (a) 20 mg / mL to 50 mg / mL of the antibody-drug conjugate, (b) 10 mM to 30 mM of histidine-histidine hydrochloride buffer, (c) 0.2 mg / mL to 0.6 mg / mL of polysorbate 80, (d) 20 mg / mL to 80 mg / mL of sucrose, wherein the pH of the reconstituted solution is 4.5 to 6.5; More preferably, the reconstituted solution comprises the following components: (a) the antibody-drug conjugate at a concentration of 20 mg / mL to 50 mg / mL, (b) about 20 mM histidine-histidine hydrochloride buffer, (c) about 0.4 mg / mL polysorbate 80, and (d) about 75 mg / mL sucrose, wherein the pH of the reconstituted solution is 5.0 to 6.

0.

16. A kit characterized in that, The invention includes a container containing a pharmaceutical composition as described in any one of claims 1 to 11, a lyophilized formulation containing an antibody-drug conjugate as described in claim 12 or 13, or a reconstituted solution containing an antibody-drug conjugate as described in claim 15.

17. Use of the pharmaceutical composition according to any one of claims 1 to 11, the lyophilized formulation containing an antibody-drug conjugate according to claim 12 or 13, the reconstituted solution containing an antibody-drug conjugate according to claim 15, or the kit according to claim 16, in the preparation of a medicament for treating and / or preventing tumors or cancer; Preferably, the tumor or cancer is a SEZ6-expressing tumor or cancer.

18. Use according to claim 17, characterized in that, The tumor or cancer mentioned is selected from lung cancer, breast cancer, liver cancer, hepatobiliary cancer, pancreatic cancer, stomach cancer, gastrointestinal cancer, intestinal cancer, colon cancer, colorectal cancer, kidney cancer, clear cell renal cell carcinoma, ovarian cancer, endometrial cancer, cervical cancer, bladder cancer, prostate cancer, testicular cancer, skin cancer, melanoma, leukemia, lymphoma, and bone cancer.

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